犬细小病毒临床样品基因组快速分离方法

Fast-separation method for genomic DNA from clinical samples of canine parvovirus

犬细小病毒是一种具有单链基因组DNA且没有囊膜的病毒,是引起幼犬出血性肠炎和心肌炎的高度接触传染性病原.研究采集疑似细小病毒感染病死犬的肠管和心脏等组织,通过组织匀浆的方法分离病毒,采用3种方法提取病毒基因组DNA.根据犬细小病毒基因序列设计特异性引物,采用PCR方法扩增病毒基因片段,琼脂糖凝胶电泳检验的扩增结果与预计结果完全一致.结果表明:CPV的分离和基因组DNA的提取方法可靠,为CPV的分离和检测及进一步研究奠定了基础.

Canine parvovirus（CPV） is a non-enveloped virus with a single-strand DNA genome and the highly contagious pathogen of hemorrhagic gastroenteritis and myocardtis in puppies.The digestive tract and the heart were obtained from the dog autopsy of the doubtful parvovirus virus infection in this investigation.Canine parvovirus is isolated by the tissue homogenation and its genomic DNA is extracted using Phenol-Chloroform,absorption of diatomite and boiling lysis technique.The primers that the partial sequence of CPV amplified by PCR were designed according to the complete sequences of CPV in Genebank of NCBI.The results of the amplified band were consisted with the predictive bands by gel electrophoresis.The result showed that the isolation of CPV and the extraction of CPV genome are the accuracy,and these methods for the isolation of CPV and the detection of CPV provide for the basis of further research.